Disequilibrium in the CD8+CD28+/CD8+CD28- T Lymphocyte Balance Is Related to Prognosis in Rats with Trinitrobenzenesulfonic Acid-Induced Colitis.

PURPOSE: The CD8+CD28+/CD8+CD28- T lymphocyte balance is vital for human ulcerative colitis (UC) but has not been defined in experimental colitis. This investigation will try to identify the changes that occur in the CD8+CD28+/CD8+CD28- T lymphocyte balance during the progression of trinitrobenzenesulfonic acid (TNBS)-induced colitis in rats. METHODS: The frequencies of blood CD8+CD28+ and CD8+CD28- T lymphocytes were detected in the rats belonging to the normal, model, and treated groups on five days using flow cytometry. The treated rats were administered with mesalazine and were euthanized after a 14-day treatment, as were the normal and model rats. The sensitivity and specificity of the CD8+CD28+/CD8+CD28- T lymphocyte balance in diagnosing early colitis were analyzed by receiver operating characteristics (ROC) curves. The frequencies of CD8+CD28+ and CD8+CD28- T lymphocytes in the colon tissue were tested via immunofluorescence. ELISA was used to measure the levels of the cytokines. Immunohistochemistry and Western blotting were used to detect the colonic expression of JAK3, STAT6, NFATc2, and GATA3. RESULTS: We found that the ratio of CD8+CD28+/CD8+CD28- T lymphocytes decreased, as did the level of interleukin-7, but not IL-12p40, IL-13, or IL-15, in the blood; however, the ratio increased along with JAK3, STAT6, NFATc2, and GATA3 in the colon of the rats with colitis. The changes were effectively reversed through the administration of mesalazine for 13 days. Surprisingly, the balance in the blood could sensitively distinguish rats with early colitis from normal rats. CONCLUSION: These data show that increase in CD8+CD28+ T cells in blood and decrease in CD8+CD28- T cells in colon are associated with experimental colitis.

[A new method for establishing a ventricular fibrillation model by TCEI in Tibetan miniature pig].

OBJECTIVE: To explore an economical, convenient, safe and efficient method for establishing a Tibetan miniature pig model of cardiac arrest (CA). METHODS: Cardiac puncture was performed in 12 Tibetan miniature pigs using two acupuncture needles. One needle was inserted into the fourth intercostal near the right side of the sternum about 3 cm in depth at an angle of 30° to 60° between the chest and the needle, and the depth was adjusted until the handle of the needle vibrated with the heartbeat without premature ventricular contraction on the electrocardiogram; the other was inserted into the subcutaneous tissue of the left armpit about 3 cm in depth without damaging important organs. The handles of the two needles were connected with 9V dry batteries to form a circuit and generate direct current stimulation. Ventricular fibrillation was produced in the pigs to induce CA by stimulation of transcutaneous electrical induction (TCEI) for 3 s, and the success rate of modeling was recorded. After an interval of 4 min without intervention, cardiopulmonary resuscitation (CPR) was performed using the standard Utstein style, and the survival of the pigs after recovery was observed. RESULTS: The success rate of ventricular fibrillation modeling was 91.67% (11/12) using this method, and CPR achieved a success rate of 45.45% (5/11) in these models. The subsequent survival of the pigs was 100% (5/5) at 24 h and 80% (4/5) at 72 h. After observation for 72 h, the resuscitated Tibetan miniature pigs were dissected, and no significant damage was found in the vital organs in the thoracic or abdominal cavities. CONCLUSIONS: We successfully established a model of CA using acupuncture needles and dry batteries in Tibetan miniature pigs, and this method is economical, convenient, safe and efficient.

CD8+CD28+/CD8+CD28- T cell equilibrium can predict the active stage for patients with inflammatory bowel disease.

BACKGROUND/AIM: The balance of blood CD8+CD28+/CD8+CD28- T cells has been verified to be vital for patients with ulcerative colitis (UC), but their role in inflammatory bowel disease (IBD) remains unknown. This investigation aimed to evaluate the efficiency of the balance in predicting the active stage in IBD patients. METHODS: Fifty-three IBD subjects, including 31 UC and 22 Crohn's disease (CD) patients, were enrolled, and their peripheral blood CD8+CD28+ and CD8+CD28- T cell levels were tested using flow cytometry. The risk factors related to prognosis were compared between UC and CD patients. A 1-year follow-up was performed for all the IBD patients, and the CD8+ T cells and their ratio were compared at the 3rd, 6th, 9th, and 12th months during follow-up. The sensitivity and specificity of the CD8+ T cell level and balance were analyzed through receiver operator characteristic (ROC) curves. The cumulative remission lasting rates (CRLRs) under the different factors were analyzed using the Kaplan-Meier method. RESULTS: Higher prescription rates of immunosuppressants, steroids, probiotics, and biological agents (BAs) were found in CD subjects in comparison to UC subjects (P=0.005, 0.024, 0.034, and 0.001), as was a higher active rate during follow-up (95.5% of CD patients vs 67.7% of UC patients, P=0.035). The CD8+CD28+ T cell level and the CD8+CD28+/CD8+CD28- T cell ratio were significantly higher in UC patients than in CD patients, but the reverse was true for CD8+CD28- T cells during follow-up at the 9th and 12th month (all P<0.05). The diagnostic models of the initial CD8+CD28+ and CD8+CD28- T cell numbers and the CD8+CD28+/CD8+CD28- T cell ratio in predicting the active stage were found to be significant, with areas under the curves (AUCs) of 0.883, 0.098, and 0.913 for UC subjects (with 95% CI: 0.709-0.940, 0.009-0.188, and 0.842-1.003; P=0.001, 0.00, and 0.000) and 0.812, 0.078, and 0.898 for CD subjects (with 95% CI: 0.683-0.957, 0.003-0.158, and 0.837-0.998; P=0.003, 0.00, and 0.000). The cut-off values showed that when the ratios were 1.30 for UC and 1.22 for CD patients, the best sensitivity and specificity were observed, with 91.6% and 89.0% for UC and 88.5% and 85.1% for CD, respectively. The CRLRs were significantly higher in female, non-BA-treated, non-surgical IBD subjects when compared to male, BA-treated, surgical subjects (P=0.031, 0.000, and 0.000). The number of CD8+CD28+ and CD8+CD28- T cells and the CD8+CD28+/CD8+CD28- T cell ratio were correlated with BA treatment and surgery (all P<0.05). CONCLUSION: The CD8+CD28+/CD8+CD28- T cell balance, expected to be a novel immunologic marker, presented a satisfactory efficiency with high sensitivity and specificity in predicting the active stage in UC and CD patients, and the balance was closely related to the use of BAs and surgery.

[Recurrence and survival analysis of postoperative patients aged 25 to 59 years with differentiated thyroid carcinoma].

OBJECTIVE: To investigate the recurrence and survival of postoperative patients with differentiated thyroid carcinoma (DTC) aged from 25 to 59 years. METHODS: We retrospectively analyzed the clinical data of 36 patients with DTC treated in our hospital from 1996 to 2011, and the recurrence and survival status of the patients were recorded. Kaplan-Meier analysis was carried out to analyze factors that affect the patient's survival. RESULTS: Nine patients died of recurrence or metastasis, and the interval between the initial surgery and recurrence ranged from 22 to 46 months. The survival time of the 36 patients ranged from 34 to 135 months with a 10-year survival rate of 75.0%. Kaplan-Meier analysis showed that male patients had a significantly shorter mean survival time than female patients (Χ2=3.164, P=0.041); the median survival time of patients aged 45-59 years was obviously shorter than that of patients aged 25-44 years (Χ2=4.622, P=0.032); the postoperative survival in patients with 131I therapy was significantly longer than those who did not receive the therapy (Χ2=4.527, P=0.033), and was not affected by total excision of the thyroid gland (Χ2=0.988, P=0.320). No significant difference was found in the median survival of patients in different clinical stages (Χ2=2.2132, P=0.167). CONCLUSION: In young and middle-aged patients with DTC, postoperative recurrence is the most likely in 2 to 4 years after the surgery. Male patients at 45-59 years of age who do not receive 131I treatment are at high risks of tumor recurrence.

[Postoperative changes of Th17/Treg balance in patients with intracranial aneurysm rupture].

OBJECTIVE: To observe the dynamic changes of Th17/Treg balance in patients following surgical intervention for intracranial aneurysm rupture. METHODS: The percentage of Th cells and the intracellular IL-17 level, Treg cell percentage and transforming growth factor -ß1 (TGF-ß1) levels were examined in 73 patients with rupture of aneurysms before and at 24 h, 72 h and 1 week after operation, with 62 patients with unruptured aneurysms and 65 healthy volunteers as the control. The correlations among the immune cells, cytokines and clinical characteristics of the patients (NIHSS, ADL and hospitalization stay) were analyzed. RESULTS: Th17 percentage and intracellular IL-17 levels were significantly higher in the patients with ruptured and unruptured aneurysms than in the healthy volunteers, and were significantly higher in patients with ruptured aneurysms than in those with unruptured aneurysms. Treg cell percentage and TGF-ß1 level were significantly lower in patients with aneurysms than in the healthy volunteers, and were lower in patients with ruptured aneurysms than in those with uruptured aneurysms (P<0.05). Patients with intracranial aneurysm rupture showed significantly increased Th17 cell percentage and IL-17 level but significantly lowered Treg cell percentage and TGF-ß1 at 24 h following the surgery (P<0.05); these changes were reversed significantly at 72 h and 1 week after the surgery. Th17 cell percentage and IL-17 level were positively correlated with NIHSS and the length of postoperative hospital stay but inversely correlated with ADL; Treg cell percentage and TGF-ß1 were inversely correlated with NIHSS and hospital stay but positively with ADL (P<0.05). CONCLUSION: In patients with intracranial aneurysms, the systemic immune inflammatory response is highlighted by excessive Th17 cells and insufficient Treg cells, which are closely related with the outcomes of the patients following surgical intervention. Evaluation of Th17/Treg balance and the cytokine levels can help to assess the prognosis of patients with aneurysm rupture.

[Immunological balance of CD8+CD28+/CD8+CD28- T lymphocytes can predict gastrointestinal hemorrhage in patients with inflammatory bowel disease].

OBJECTIVE: To evaluate the sensitivity and specificity of CD8+CD28+/CD8+CD28- T lymphocyte balance in predicting the gastrointestinal hemorrhage (GH) in patients with inflammatory bowel disease (IBD). METHODS: Forty-nine IBD patients, including 30 with ulcerous colitis (UC) and 19 with Crohn's disease (CD), were enrolled to test peripheral blood CD8+CD28+ and CD8+CD28- T cells using flow cytometry. All the patients were followed up for one year. The receiver-operating characteristic (ROC) curves were used to test the efficiency of CD8+CD28+/CD8+CD28- T lymphocyte balance to predict GH. The differences in lasting time of remission (LTR) under different factors were compared using Kaplan-Meier survival analysis, and the correlation between CD8+ T lymphocytes and the factors were analyzed. RESULTS: The utilization rates of immunosuppressant, steroids, and biological agent (BA) were significantly higher in CD patients than in UC patients (P=0.003, 0.043 and 0.002, respectively). The frequencies of CD8+CD28+T cells were obviously higher in UC patients than those in CD patients (t=3.022, P=0.004). CD8+CD28+T cells, CD8+CD28- T cells, and especially CD8+CD28+/CD8+CD28- ratio (area under curve of 0.977, P=0.000; cut-off value of 1.14 [13.95%/12.24%] with a sensitivity of 93.3% and a specificity of 91.2%) showed good efficiencies in predicting GH (P<0.01). The mean and median of LTR of IBD patients who did not receive BA or surgical treatment were significantly longer (Χ2=9.730, P=0.002; Χ2=15.981, P=0.000). CD8+CD28+/CD8+CD28- ratio was significantly related to both BA (P=0.009) and surgery (P=0.038). CONCLUSION: Both decreased CD8+CD28+T cells and elevated CD8+CD28-T cells are closely correlated with GH, and their ratio can predict the occurrence of GH with a high sensitivity and specificity and is correlated with BA and surgery at the cut-off value of 1.14.